Stemi 2000 c

The transformants generated and wildtype reference strain IPO323 were spotted (1.5 μl; 1×10⁸ spores/ml) onto N-deprivation solid medium and water agar. Colonies were analysed stereomicroscopically using binocular Zeiss Stemi 2000-C (Carl Zeiss Microscopy GmbH) after 7, 14 and 21 days of incubation at 18°C to monitor the dynamics of dimorphic switch of strains generated. Morphology scoring was based on macroscopic colony appearance at 14 days, with consideration of appearance at 21 days in questionable cases. The mutants were considered as dimorphic switch deficient when their growth and switching inability was comparable with that of ΔZthog1 mutant strain as reference after 7 days post inoculation of N-deprivation and water agar medium.

The main characters Broun [10 –13 ] used to delineate species were body size, body colour, pronotum shape, pronotum size, density of pronotal punctures, and density of elytral punctures. Shaw [15 ], whose study was based almost entirely on reexamination of Broun’s specimens at the British Museum of Natural History (BMNH), primarily used external shape and pronotal puncture density. However, neither Broun nor Shaw quantified the variation in morphology within and between putative species, but rather used relative estimates to indicate variation. It is clear that some of the characters, such as body colour, shape and size of pronotal punctures can vary greatly within populations (P.Doddala, pers. obs.).
In this study, we quantified a set of the external morphological characters previously used (Fig 2A). External morphological characters were recorded from randomly selected individuals from each locality / sample using a digital camera (Moticam 2000 2.0 MP USB 2.0; Motic Group Co., Ltd.) fitted to a dissecting microscope (Zeiss Stemi 2000-c; Carl Zeiss, Inc.). Motic Images Plus v.2.0 (Motic Group Co., Ltd.) was used to record measurements from those images.

Preliminary experiments showed that the shoot apices did not develop during the 3-week cold hardening period, therefore the initial development of flower primordia was monitored at the end of the 3-week recovery period, when the apices were isolated from the crowns of the seedlings under a Zeiss Stemi 2000-C stereomicroscope (Carl Zeiss Mikroskopie, Jena, Germany). The photos were taken with a Camedia digital camera using standardized exposure times and sensor settings. The photos of the apices are shown in Figure 8 and in Additional file 5. The developmental stages of the apices were determined based on the scale of Gardner et al. [51 (link)], which takes into account the appearance of new structures. The scale between 0 and 8 corresponds to the following developmental stages: 0 – vegetative apex, 1 – early elongation of the apex, 2 – elongation with single ridge, 3 – double ridge indicating the vegetative/generative transition, 4 – enlargement of spikelet primordia, 5 – empty glume primordia, 6 – lemma glume primordia, 7 – floret and anther primordia, 8 – terminal spike.

Pictures of the mini-implants were obtained under optical microscope (Zeiss Stemi 2000-C, Zeiss) observed at 1.6 x magnification. The pictures of the surfaces were analyzed with Axio Vision program (Zeiss) which were made of linear and angular measurements, as shown in Figures 2 and 3.
Figure 2Figure 3

The quality of each suppository was visually inspected directly after the manufacturing process. Photographic images were exemplarily taken, and microscopic imaging was obtained using a reflected light microscope (Zeiss Stemi 2000-C with Zeiss CL 1500 ECO, AxioCam ICc 1 and AxioVision software v4.8.2, all Carl Zeiss AG, Oberkochen, Germany). Further imaging of the suppository morphology was performed for PEG formulations by scanning electron microscopy (SEM; Phenom FP 3950/00, L.O.T.-Oriel Laser Optik Technologie GmbH und Co. KG, Darmstadt, Germany) with an acceleration voltage of 5 kV. Prior to the SEM examination, the samples were coated with gold–palladium under vacuum using a mini sputter coater (SC7620, Quorum Technologies Ltd., East Sussex, UK).
Furthermore, the printing process for samples of each suppository design was terminated at a height of 15 mm to obtain insights into the internal structure of the suppositories without the necessity to destroy the suppositories, which may be associated with structural changes.

Zooplankton samples were collected by vertical towing a plankton net (70 μm mesh) from the bottom to the water surface or by filtering through a net, the water being collected with a measuring bucket. The samples were concentrated to 100 ml and fixed to a final concentration of 4% formalin solution (Mordukhay-Boltovskoy 1975 ). Zooplankton specimens were examined using a Zeiss Stemi 2000C stereomicroscope (Carl Zeiss Microscopy, Germany) and a detailed morphological analysis was performed using an Olympus CX43 light microscope (Olympus Crp., Japan). Studies were conducted mostly in the lower reaches and estuaries of the rivers. A total of 200 zooplankton samples were processed. After processing, all samples were stored in the authors' personal collections for further detailed taxonomic and morphological studies.